ArticleMolecular and biochemical parasitology
A unified approach towards Trypanosoma brucei functional genomics using Gibson assembly.
Article in Molecular and biochemical parasitology. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 15 citations in OpenAlex.
- Myosin F controls actin organization and dynamics inMolecular biology of the cell · 2024Article
- Revealing spatio-temporal dynamics with long-term trypanosomatid live-cell imaging.PLoS pathogens · 2022Article
- The Trypanosoma brucei subpellicular microtubule array is organized into functionally discrete subdomains defined by microtubule associated proteins.PLoS pathogens · 2021Article
- Article
- Branched late-steps of the cytosolic iron-sulphur cluster assembly machinery of Trypanosoma brucei.PLoS pathogens · 2018Article
- Identification of TOEFAZ1-interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis.Molecular microbiology · 2018Article
- TbSmee1 regulates hook complex morphology and the rate of flagellar pocket uptake in Trypanosoma brucei.Molecular microbiology · 2018Article
- Simplified inducible system for Trypanosoma brucei.PloS one · 2018Article
- A functional analysis of TOEFAZ1 uncovers protein domains essential for cytokinesis inJournal of cell science · 2017Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Trypanosoma brucei is the causative agent of human African trypanosomiasis and nagana in cattle. Recent advances in high throughput phenotypic and interaction screens have identified a wealth of novel candidate proteins for diverse functions such as drug resistance, life cycle progression, and cytoskeletal biogenesis. Characterization of these proteins will allow a more mechanistic understanding of the biology of this important pathogen and could identify novel drug targets. However, methods for rapidly validating and prioritizing these potential targets are still being developed. While gene tagging via homologous recombination and RNA interference are available in T. brucei, a general strategy for creating the most effective constructs for these approaches is lacking. Here, we adapt Gibson assembly, a one-step isothermal process that rapidly assembles multiple DNA segments in a single reaction, to create endogenous tagging, overexpression, and long hairpin RNAi constructs that are compatible with well-established T. brucei vectors. The generality of the Gibson approach has several advantages over current methodologies and substantially increases the speed and ease with which these constructs can be assembled.
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